Optimized transitory ectopic expression of promastigote surface antigen protein in Nicotiana benthamiana, a potential anti-leishmaniasis vaccine candidate

Fasciola gigantica diagnosis usually performed by detection of worm eggs presence in the feces,but this conventional method has many disadvantages. Early diagnosis (early detection) cannot be performed in conventional methods because the worms in the host's body began to lay eggs at the age of 8–12 weeks of patency. The current detection method that is based on antibodyantigen reactions using excreted/secreted (ES) liquid by adult F. gigantica, is believed to be used for the early detection of fasciolosis. This study aimed to characterize the antigenic components of F. gigantica extretory/secretory products that could be used as a vaccine candidate development for early fasciolosis diagnostics. ES products were separated by PEG4000 at various concentrations (8%, 16%, 24%), then precipitates (pellets) obtained were dialyzed and characterized using SDSPAGE and Western blotting. Results from SDSPAGE showed that there were 18 proteins bands with 7–70 kDa molecular weights. Western blotting on pellets derived from PEG separation at various concentrations affirmed that the proteins of 50, 25 and 20 kDa were antigenic at 8% PEG concentration, the 25 kDa and 50 kDa were antigenic at 16% PEG concentrations and the 25 kDa was antigenic at 25% PEG concentration

Download Full-text

Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in Nicotiana benthamiana Plant as a Vaccine Candidate against COVID-19

Vaccines ◽

10.3390/vaccines9111337 ◽

2021 ◽

Vol 9 (11) ◽

pp. 1337

Author(s):

Tarlan Mamedov ◽

Damla Yuksel ◽

Merve Ilgın ◽

Irem Gürbüzaslan ◽

Burcu Gulec ◽

...

Keyword(s):

Nicotiana Benthamiana ◽

Vaccine Candidate ◽

Plant Biomass ◽

Polyclonal Antibodies ◽

High Titer ◽

Benthamiana Plant ◽

Global Public Health ◽

Neutralizing Activity ◽

N Protein ◽

The World

The COVID-19 pandemic has put global public health at high risk, rapidly spreading around the world. Although several COVID-19 vaccines are available for mass immunization, the world still urgently needs highly effective, reliable, cost-effective, and safe SARS-CoV-2 coronavirus vaccines, as well as antiviral and therapeutic drugs, to control the COVID-19 pandemic given the emerging variant strains of the virus. Recently, we successfully produced receptor-binding domain (RBD) variants in the Nicotiana benthamiana plant as promising vaccine candidates against COVID-19 and demonstrated that mice immunized with these antigens elicited a high titer of RBD-specific antibodies with potent neutralizing activity against SARS-CoV-2. In this study, we engineered the nucleocapsid (N) protein and co-expressed it with RBD of SARS-CoV-2 in Nicotiana benthamiana plant to produce an antigen cocktail. The purification yields were about 22 or 24 mg of pure protein/kg of plant biomass for N or N+RBD antigens, respectively. The purified plant produced N protein was recognized by N protein-specific monoclonal and polyclonal antibodies demonstrating specific reactivity of mAb to plant-produced N protein. In this study, for the first time, we report the co-expression of RBD with N protein to produce a cocktail antigen of SARS-CoV-2, which elicited high-titer antibodies with potent neutralizing activity against SARS-CoV-2. Thus, obtained data support that a plant-produced antigen cocktail, developed in this study, is a promising vaccine candidate against COVID-19.

Download Full-text

A highly pathogenic porcine reproductive and respiratory syndrome virus candidate vaccine based on Japanese encephalitis virus replicon system

PeerJ ◽

10.7717/peerj.3514 ◽

2017 ◽

Vol 5 ◽

pp. e3514 ◽

Cited By ~ 1

Author(s):

Pingsheng Hu ◽

Xiaoming Chen ◽

Lihong Huang ◽

Shukai Liu ◽

Fuyu Zang ◽

...

Keyword(s):

Japanese Encephalitis Virus ◽

Japanese Encephalitis ◽

Vaccine Candidate ◽

Expression System ◽

Ectopic Expression ◽

Encephalitis Virus ◽

Economic Losses ◽

Respiratory Syndrome Virus ◽

Response Analysis ◽

Effective Prevention

In the swine industry, porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease which causes heavy economic losses worldwide. Effective prevention and disease control is an important issue. In this study, we described the construction of a Japanese encephalitis virus (JEV) DNA-based replicon with a cytomegalovirus (CMV) promoter based on the genome of Japanese encephalitis live vaccine virus SA14-14-2, which is capable of offering a potentially novel way to develop and produce vaccines against a major pathogen of global health. This JEV DNA-based replicon contains a large deletion in the structural genes (C-prM-E). A PRRSV GP5/M was inserted into the deletion position of JEV DNA-based replicons to develop a chimeric replicon vaccine candidate for PRRSV. The results showed that BALB/c mice models with the replicon vaccines pJEV-REP-G-2A-M-IRES and pJEV-REP-G-2A-M stimulated antibody responses and induced a cellular immune response. Analysis of ELSA data showed that vaccination with the replicon vaccine expressing GP5/M induced a better antibodies response than traditional DNA vaccines. Therefore, the results suggested that this ectopic expression system based on JEV DNA-based replicons may represent a useful molecular platform for various biological applications, and the JEV DNA-based replicons expressing GP5/M can be further developed into a novel, safe vaccine candidate for PRRS.

Download Full-text